Rapid Detection of Panax ginseng by Loop-Mediated Isothermal Amplification and Its Application to Authentication of Ginseng

Abstract

We have developed a novel method called loop-mediated isothermal amplification (LAMP) to detect Panax ginseng, the botanical source of Ginseng (Ginseng Radix), and to distinguish P. ginseng from Panax japonicus. Six allele-specific primers (two outer primers, two inner primers, and two loop primers) were designed based on the 18S ribosomal RNA gene sequence of P. ginseng, and LAMP was performed using those primers and total DNA extracted from P. ginseng as template. Amplifications were observed from approximately 30 min onwards at DNA concentrations of 0.5 to 10.0 ng. The presence of loop primers shortened the reaction time considerably. In contrast, in the reactions using total DNA from P. japonicus as template, no amplifications were observed. LAMP also enabled us to distinguish Ginseng from Japanese Ginseng (Panacis Japonici Rhizoma). LAMP was proven to be a rapid, highly sensitive, and specific method for the detection of P. ginseng and Ginseng.

Identification of Baiying (Herba Solani Lyrati) commodity and its toxic substitute Xungufeng (Herba Aristolochiae Mollissimae) using DNA barcoding and chemical profiling techniques

“…In this study, we have established five DNA barcodes that could be served as novel molecular markers for the authentication of Baiying and Xungufeng. The polymorphic sites could be used to design specific primers for the development of efficient molecular authentication techniques, including multiplex polymerase chain reaction and novel isothermal amplification (Jigden et al, 2010;Sasaki, Komatsu, & Nagumo, 2008). TLC and HPLC profiles of Baiying and Xungufeng provided alternative authentication methods using chemical approach, while the cytotoxicities of these two herbs could be potential biological markers for identification.…”

Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of <i>Cannabis sativa</i>

“…19,20) It does not need any special devices or experienced technicians; thus, it is widely used for various purposes, such as detecting viruses and medicinal herbs. [21][22][23][24] One of the most species-specific genes in C. sativa is the THCA synthase gene that consists of a 1632 bp open reading frame. THCA synthase gene was cloned and polymorphism analysis revealed that 63 nucleotide substitutions divided the species into two chemotypes: the THCA-rich type (drug type), such as marijuana, and the THCA-poor type (fiber type), such as hemp.…”

Molecular Authentication of Magnoliae Flos Using Robust SNP Marker Base on trnL-F and ndhF Region

“…Several molecular methods, such as arbitrarily primed polymerase chain reaction (AP-PCR) (Cheung et al, 1994), random amplified polymorphic DNA (RAPD) (Cui et al, 2003;Shim et al, 2003;Shaw and But, 1995), loop-mediated isothermal amplification (LAMP) (Sasaki et al, 2008), sequence characterized-amplified region (SCAR) (Choi et al, 2008;Wang et al, 2001), restriction fragment length polymorphism (RFLP) (Ngan et al, 1999), amplification fragment length polymorphism (AFLP) (Ha et al, 2002), and DNA microarray (Zhu et al, 2008) have previously been described. The main drawback of RAPD are lack of reliability and reproducibility.…”